Aminoquanidine carboxylates for the treatment of obesity

ABSTRACT

The present invention provides novel compounds of the formulae (I): AG--(CH 2 ) n  --CO 2  R 1  and (II) or pharmacologically acceptable salts thereof, wherein AG is: a) N-aminoguanidine, b) N,N&#39;-diaminoguanidine, or c) N,N&#39;,N&#34;-triaminoguanidine; and n, R 1  and R 2  have the meanings given in the description as well as a new use for the compounds of formula (III) wherein R 3  has the meanings given in the description for the treatment of non-insulin dependent diabetes mellitus (NIDDM).

CROSS REFERENCE TO RELATED APPLICATIONS

This application is the national phase of international applicationPCT/US95/14126, filed Nov. 13, 1995; which is a continuation of U.S.Ser. No. 08/484,547, filed Jun. 7, 1995; which is a continuation-in-partapplication of U.S. Ser. No. 08/344,274, filed Nov. 23, 1994, nowabandoned.

The present invention provides novel compounds and a novel method fortreating: non-insulin dependent diabetes mellitus (NIDDM); diabeticcomplications resulting from excessive non-enzymatic glycosylation ofproteins in non-insulin dependent and insulin-dependent diabetesmellitus; impaired glucose tolerance; and obesity.

BACKGROUND OF THE INVENTION

Non-insulin dependent diabetes mellitus, or NIDDM, and Type II diabetesare synonymous. NIDDM patients have an abnormally high blood glucoseconcentration when fasting and delayed cellular uptake of glucosefollowing meals or after a diagnostic test known as the glucosetolerance test. NIDDM is diagnosed based on recognized criteria(American Diabetes Association, Physician's Guide to Insulin-Dependent(Type I) Diabetes, 1988; American Diabetes Association, Physician'sGuide to Non-Insulin-Dependent (Type II) Diabetes, 1988).

Insulin-Dependent diabetes mellitus, IDDM, and Type I diabetes aresynonymous. IDDM patients have an abnormally high blood glucoseconcentration when fasting and delayed cellular uptake of glucosefollowing meals or after a diagnostic test known as the glucosetolerance test. IDDM is diagnosed based on recognized criteria (AmericanDiabetes Association, Physician's Guide to Insulin-Dependent (Type I)Diabetes, 1988).

Impaired glucose tolerance occurs when the rate of metabolic clearanceof glucose from the blood is less than that commonly occurring in thegeneral population after a standard dose of glucose has been orally orparenterally administered (American Diabetes Association, Physician'sGuide to Non-Insulin-Dependent (Type II) Diabetes, 1988). Impairedglucose tolerance can occur in NIDDM, IDDM, gestational diabetes andobesity. Impaired glucose tolerance can also occur in individuals notmeeting the diagnostic criteria for these disease states. Impairedglucose tolerance in non-diabetic individuals is a predisposing factorfor the development of NIDDM.

Obesity is a condition in which there is an increase in body fat contentresulting in excess body weight above the accepted norms for age,gender, height, and body build (Bray, Obesity, An Endocrine Perspective,p. 2303, Multihormonal Systems and Disorders (1989)). Accepted normshave been determined by life insurance mortality experience and byincidence of morbidity in relation to body composition. The excessmortality that occurs in obese individuals results from diseases thatare predisposed by this condition. They include cancer, cardiovasculardisease, digestive disease, respiratory disease and diabetes mellitus.

In patients with chronic hyperglycemia such as occurs in non-insulindependent diabetes and insulin-dependent diabetes, glucose-dependentprotein crosslinking occurs at a rate in excess of the norm (Bunn,American Journal of Medicine, Vol. 70, p. 325, 1981) resulting inaltered tertiary protein structure (Brownlee, Chapter 18, DiabetesMellitus, p. 279, 1990). Excessive non-enzymatic glycosylation ofproteins contributes to diabetic complications and complications ofaging in non-diabetic humans, such as neuropathy, nephropathy,retinopathy, hypertension, and atherosclerosis (Brownlee, 1990, supra).

Hyperglycemia is defined as blood glucose concentration in excess of theaccepted norm for the general population (American Diabetes Association,Physician's Guide to Non-Insulin-Dependent (Type II) Diabetes, 1988).

While the relationship between these conditions is known, it would be anadvantage to have a drug which can treat or prevent all of them.

INFORMATION DISCLOSURE

3-(1-(Aminomethyl)hydrazino)) propanoic acid is reported in JP 54128523(Chem. Abstr. 92:75899h) to be a fungicide and insecticide. Thesynthesis of N-(hydrazinoiminomethyl)-glycine is reported in: Gante, J.Chem. Ber. 1968, 101, 1195. Certain alkylide-amino guanidine derivativesare described in U.S. Pat. No. 5,272,165 titled "Inhibiting advancedglycosylation of body proteins--using 2-alkylidene-amino:guanidinederiv., used e.g. for treating diabetic side-effects or esp. preventingtooth staining." Aminoguanidine analogs of arginine are disclosed in DE4244539-A1 and WO 9104-023-A. U.S. Pat. No. 5,132,453 discloses thatN6-(hydrazino:imino:-methyl)-lysine is useful as an inhibitor of nitricoxide formation and for treating hypertension. EP-230-037-A disclosescertain new 2-substituted-guanidine derivatives having antiischaemic andcardioprotective activity. U.S. Pat. No. 3,412,105 disclosesβ-Aryl-N-guanidino-(β-alanines or α-carboxy-β-alanines) as MAOinhibitors and long acting hypotensives.

SUMMARY OF THE INVENTION

The present invention particularly provides:

(1) A compound of the formulae I or II: ##STR2## or a pharmacologicallyacceptable salt thereof, wherein AG is

a) N-aminoguanidine,

b) N,N'-diaminoguanidine, or

c) N,N',N"-triaminoguanidine;

wherein n is an integer from 1-5;

wherein R¹ is

a) hydrogen,

b) phenyl,

c) C₁ -C₅ alkyl, or

d) C₁ -C₃ alkyl-phenyl; and

wherein R² is

a) hydrogen,

b) phenyl,

c) C₁ -C₁₀ alkyl, or

d) C₁ -C₅ alkyl-phenyl

with the following provisos:

a) in Formula I, when n is 2, R₁ is other than hydrogen;

b) in Formula I, when n is one, R₁ is other than methyl;

c) in Formula II, when R₂ is ethyl, R₁ is other than hydrogen;

d) in Formula II, when R₂ is phenyl, R₁ is other than hydrogen; and

e) in Formula I, when n is 3, R₁ is other than hydrogen.

(2) a method for treating or preventing non-insulin dependent diabetesmellitus in a patient suscepible to or experiencing said NIDDMcomprising the systemic administration of an amount effective to treator prevent NIDDM of a compound of the formula III ##STR3## wherein R³ ishydrogen, methyl, ethyl, CH₂ phenyl, or n-hexyl.

For the generic formulae I and II, attachment of the AG fragment isunspecified, i.e. bonding to the adjacent carbon may occur at any one ofthe nitrogens of the AG fragment. The remaining nitrogens of the AGfragment are unsubstituted.

The carbon atom content of the carbon containing moieties is indicatedby a prefix "C_(i) -C_(j) " wherein i is the lowest number of carbonatoms and j is the highest number of carbon atoms.

Examples of alkyl groups having from 1 to 10 carbon atoms include, forexample, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl,n-pentyl, isoamyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, andother isomeric forms thereof.

Examples of pharmaceutically acceptable acid addition salts include:acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate,bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate,hexanoate, hydrochloride, hydrobromide, hydroiodide,2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate,2-naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate,persulfate, 3-phenylpropionate, picrate, pivalate, propionate,succinate, tartrate, thiocyanate, tosylate, and undecanoate.

The dose of compounds of formula I-III to be used is between 0.1 and 100mg/kg body weight daily. The preferred dose is 1-50 mg/kg/day.Administration may be by oral, parenteral, intranasal, buccal,sublingual, intrarectal, or transdermal routes. The oral route ispreferred.

Novel compounds of the invention are given by the generic formulae I andII. Known compounds claimed for use in the treatment of NIDDM arerepresented by formula III.

Of the compounds of this invention, represented by generic formulae Iand II, the compounds listed in Table 1 are especially preferred andtheir preferred utility is in the treatment of NIDDM and itscomplications.

Table 2 contains a list of related compounds which are not claimed. Theyare included to demonstrate the surprising effect of the claimedcompounds by showing that these compounds, which are closely related tothe claimed compounds, are not considered active at the highest dosetested.

Table 3 contains a list of compounds within the generic scope embodiedin the generic formulae I and II which failed to exhibit activity at thehighest dose tested and thus constitute exceptions, as seen by theprovisos in claim 1.

Table 4 contains a list of novel compounds specifically claimed withinthe invention. Procedures for their preparation are given in Section 4.

Table 5 contains a list of known compounds being claimed for use in thetreatment of NIDDM.

Thus, the present invention provides novel and known compounds havingsurprising and unexpected antidiabetic properties.

Administration of the compounds of this invention to KKAy mice at a doseof approximately 100-500 mg/kg/day results in the partial or completeamelioration of hyperglycemia in this rodent model of non-insulindependent diabetes mellitus (Specific compounds are listed in Tables 4and 5; see Chang, Wyse, Copeland, Peterson, and Ledbetter, Diabetes1985, p. 466, 1986). KKAy mice are insulin resistant (Chang, et al,supra) and the finding that the non-fasting blood glucose level isreduced in these animals indicates that insulin resistance is mostprobably less after treatment with the claimed compounds. KKAy mice areobese compared to normal, outbred mice (Chang, et al, supra) andadministration of compounds of the invention results in weight loss.

Administration of N-(dihydrazinomethylene)-glycine, the preferredcompound in this series, to diabetic KKAy mice for 4 days decreased thenon-fasting blood glucose level of the animals (see Table 6). A dose of60 mg/kg/day produced a 35% decrease in the blood glucose level that wasstatistically significant compared to the control. Higher doses producedstill greater reductions in the blood glucose concentration.3-Guanidinopropionic acid at 500 mg/kg/day produced an approximatelysimilar reduction in blood glucose concentration as was achieved with 60mg/kg/day of the N-(dihydrazinomethylene)-glycine.

Administration of N-(dihydrazinomethylene)-glycine to diabetic KKAy micefor 4 days decreased the body weight of the animals (see Table 6). Adose of 100 mg/kg/day produced a 4% decrease in the body weight that wasstatistically significant compared to the control. Higher doses produceda still greater reduction in the excess body weight of KKAy mice.3-Guanidinopropionic acid at 500 mg/kg/day produced an approximatelysimilar reduction in the body weight of KKAy mice as was achieved with100 mg/kg/day of the N-(dihydrazinomethylene)-glycine.

Administration of N-(dihydrazinomethylene)-glycine to normal C57BL miceat 100 mg/kg decreased the fasting blood glucose concentration of theseanimals (Table 7).

Administration of N-(dihydrazinomethylene)-glycine to diabetic KKAy miceor normal C57BL mice at 100 mg/kg results in improved glucose toleranceas shown by lower blood glucose levels after injection of a standardtest dose of glucose (Table 7).

Non-enzymatic glycosylation of proteins is the initial step inglucose-dependent crosslinking of proteins (Brownlee, supra).Non-enzymatic glycosylation of human serum albumin is reduced byN-(dihydrazinomethylene)-glycine, N-(hydrazinoiminomethyl)-glycine, and[2-(aminoiminomethyl)hydrazino]-, monohydrochloride acetic acid in vitro(Table 8). Aminoguanidine, which has previously been shown to inhibitnon-enzymatic glycosylation of proteins in vitro (Khatami, Suldan,David, Li, and Rockey, Life Sciences, vol. 43, p. 1725-1731, 1988) andin vivo (Brownlee, supra), is also effective in this assay (Table 8).3-Guanidinopropionic acid had no effect on non-enzymatic glycosylationof albumin in this assay.

In patients with diabetes mellitus, there are several metabolicdisorders that would be of therapeutic benefit to correct: theabnormally elevated blood level of glucose in the fed and fasted states,the delayed clearance of glucose from the blood stream (AmericanDiabetes Association, Physician's Guide to Insulin-Dependent (Type I)Diabetes, 1988; American Diabetes Association, Physician's Guide toNon-Insulin-Dependent (Type II) Diabetes, 1988), and the excessiveglycosylation of proteins which contributes to the development ofdiabetic complications (Brownlee, supra). Furthermore, obesity isfrequently associated with non-insulin dependent diabetes mellitus andaggravates the disordered glucose metabolism in these patients (Hortonand Jeanrenaud, Chapter 27, Obesity and Diabetes Mellitus, 1990). Theoptimal treatment for non-insulin dependent diabetes mellitus wouldcorrect all of these disorders. Excessive glycosylation of proteins,such as can occur in non-insulin dependent diabetes mellitus andinsulin-dependent diabetes mellitus patients, can be prevented byblocking the chemical reaction of glucose and protein molecules(Brownlee, supra) and reducing the abnormal elevation of blood glucoseconcentration in the diabetic state (Holman and Turner, DiabeticMedicine, 5:582-588, 1988; Benjamin and Sacks, Clin Chem., 4015:683-687,1994). The most desirable treatment would act by both methods so as tomore completely reduce the rate of non-enzymatic protein glycosylation.

It is the ability of the claimed compounds to positively effect multiplemetabolic defects comprising diabetes mellitus and to prevent metabolicdefects by more than one mechanism that clearly distinguishes theirpharmacologic actions from other guanidine compounds that havepreviously been claimed as treatments for diabetes mellitus. The claimedcompounds are unexpectedly superior to aminoguanidine, diaminoguanidine,3-guanidinopropionic acid, and metformin in the treatment of NIDDMbecause they offer a more complete spectrum of desirable activities andare effective in lower doses.

The claimed compounds offer unexpected advantages in the treatment ofdiabetes mellitus compared to diaminoguanidine and aminoguanidine sincethe claimed compounds act metabolically to reduce excessive bloodglucose concentration as well as directly blocking non-enzymaticglycosylation of proteins. The claimed compounds are unexpectedlysuperior to aminoguanidine and diaminoguanidine in the treatment ofimpaired glucose tolerance or obesity since aminoguanidine anddiaminoguanidine lack efficacy in this regard. Aminoguanidine anddiaminoguanidine inhibit non-enzymatic glycosylation of proteins invitro and the formation of advanced glycosylation endproducts in vivo(Kumari, Umar, Bansal, and Sahib, Diabetes, 40:1079-1084, 1991). Basedon its inhibition of non-enzymatic protein glycosylation, aminoguanidinehas been suggested to have utility in the treatment of diabetes(Brownlee, supra). Aminoguanidine has no effect on the blood glucoselevel of normal rodents or rats made diabetic by injection of alloxan orstreptozotocin (Kumari, Umar, Bansal, Sahib, supra; Yagihashi, Kamijo,Baba, Yagihashi, and Nagai, Diabetes, 41:47-52, 1992; Edelstein andBrownlee, Diabetologia, 35:96-97, 1992; Oxlund and Andreassen,Diabeterologia, 35:19-25, 1992). Diaminoguanidine has no effect on theblood glucose level of normal or alloxan-diabetic rats (Kumari, Umar,Bansal, Sahib, supra). Aminoguanidine has no effect on the body weightof normal or diabetic rats (Kumari, Umar, Bansal, Sahib, supra;Yagihashi, Kamijo, Baba, Yagihashi, and Nagai, supra; Oxlund andAndreassen, Diabetologia, 35:19-25, 1992) or results in an increase inbody weight of human and rats (Baylin, Horakova, and Beaven,Experientia, 31:562, 1975). Diaminoguanidine does not affect the bodyweight of normal or alloxan-diabetic rats (Kumari, Umar, Bansal, Sahib,supra). An effect by aminoguanidine or diaminoguanidine on glucosetolerance has yet to be demonstrated.

The claimed compounds are unexpectedly superior to 3-guanidinopropionicacid in the treatment of diabetes mellitus since the latter is lesspotent in the control of hyperglycemia and lacks the ability to inhibitthe non-enzymatic glycosylation of proteins. The claimed compounds areunexpectedly superior to 3-guanidinopropionic acid in the treatment ofimpaired glucose tolerance or obesity because of the greater potency ofthese compounds. 3-Guanidinopropionic acid has previously been shown toreduce hyperglycemia and excess body weight and to improve glucosetolerance in diabetic rodents (Meglasson, Wilson, Yu, Robinson, Wyse,and de Souza, J. Pharm. and Exp. Therapeutics, 266:1454-1462, 1993). Thepreferred compound in this claim, N-(dihydrazinomethylene)-glycine, ismore potent than 3-guanidinopropionic acid in reducing the abnormallyelevated blood glucose level and body weight of KKAy mice. To reducedthe blood glucose level of KKAy mice by 20% required 130 mg/kg/day ofthe latter compound. A similar reduction in the blood glucose levelcould be achieved with a dose of 30 mg/kg/day ofN-(dihydrazinomethylene)-glycine. N-(dihydrazinomethylene)-glycineadministered to KKAy mice at 60 mg/kg/day was approximately as effectiveas 500 mg/kg/day of 3-guanidinopropionic acid. 3-Guanidinopropionic acidimproves glucose tolerance in diabetic KKAy mice when administered inthe chow as a 1% admixture which would deliver a dose of approximately1000 mg/kg/day (U.S. Pat. No. 5,132,324). By comparison,N-(dihydrazinomethylene)-glycine improved the glucose tolerance ofnormal C57BL and diabetic KKAy mice when administered at 100 mg/kg/day.With respect to reducing body weight, 100 mg/kg/day ofN-(dihydrazinomethylene)-glycine was approximately as effective as 500mg/kg/day of 3-guanidinopropionic acid. 3-Guanidinopropionic acid doesnot inhibit non-enzymatic glycosylation of albumin in vitro in contrastto the claimed compounds.

The claimed compounds are unexpectedly superior to metformin in thetreatment of diabetes mellitus, glucose intolerance, and obesity sincethe latter is less potent when tested in the same animal model as theclaimed compounds. Also, with respect to its efficacy in reducing bodyweight and preventing non-enzymatic protein glycosylation, the discloseddata for metformin are contradictory and do not reveal a consistentresult. Metformin has previously been shown to reduce hyperglycemia innon-insulin dependent diabetic patients when administered at 1000-3000mg/day and to increase the rate of glucose clearance in such patientswhen administered at 1500-2500 mg/day (Bailey, Diabetes Care,15:755-772, 1992). Rodents are less sensitive to metformin than humansand therefore higher doses (based on body weight) are required todemonstrate glycemic effects (Bailey, Flatt, Wilcock, and Day, Frontiersin Diabetes Research, pp. 277-282, 1990; Penicaud, Hitier, Ferre, andGirard, Biochem. J. 262:881-885, 1989). Chronic oral administration ofmetformin reduces hyperglycemia when administered to neonatalstreptozotocin-diabetic rats at 100 mg/kg/day (Rossetti, DeFronzo,Gherzi, Stein, et al, Metabolism, 39:425-435, 1990), to DBM mice at 400mg/kg/day (Bailey, Flatt, Wilcock, and Day, supra), to Zucker fa/fa ratsat 350 mg/kg/day (Penicaud, Hitier, Ferre, and Girard, supra), and toKKAy mice at 300 mg/kg/day or more (Meglasson, Wilson, Yu, Robinson, deSouza, supra). Chronic oral administration of metformin did not affectthe blood glucose concentration in normal mice receiving 250 mg/kg/day,in streptozotocin-diabetic mice receiving 250 mg/kg/day (Bailey, Flatt,Wilcock, and Day, supra), or diabetic ob/ob mice receiving 250 mg/kg/day(Bailey, Flatt, and Ewan, Arch. Int. Pharmacodyn., 282:233-239, 1986).Acute administration of 264 mg/kg metformin or its analog buformin at132 mg/kg did not affect the blood glucose level of rats (Tutwiler andBridi, Diabetes, 27:868-876, 1978). When the preferred compound in thisclaim, N-(dihydrazinomethylene)-glycine was tested in KKAy mice it wasmore potent than metformin in reducing the abnormally elevated bloodglucose level in this model. To reduce the blood glucose level of KKAymice by 25% required 300 mg/kg/day of metformin (Meglasson, Wilson, Yu,Robinson, Wyse, and de Souza, supra). A similar reduction in the bloodglucose level could be achieved with a dose of 30-60 mg/kg/day ofN-(dihydrazinomethylene)-glycine. With respect to increasing glucosetolerance metformin has been reported to not affect glucose tolerance innormal rats when given at a dose of 750 mg/kg (Tutwiler and Bridi,supra) or in normal mice when given at 50 mg/kg (Bailey, Flatt, Wilcock,and Day, supra). When given to normal mice or streptozotocin-diabeticrats at 250 mg/kg oral glucose tolerance was increased (Bailey, Flatt,Wilcock, and Day, supra). By comparison,N-(dihydrazinomethylene)-glycine increased glucose tolerance whenadministered to normal C57BL or diabetic KKAy mice at a lower dose, 100mg/kg. With respect to reducing body weight, metformin has been reportedto cause weight loss in non-insulin dependent diabetic patients treatedfor one year (Bailey, supra) or to have no significant effect on thebody weight of obese non-insulin dependent diabetic patients treated fora similar length of time (Multi-centre Study, Diabetologia, 24:404-411,1983). Metformin did not cause weight loss in diabetic ob/ob mice whenadministered at 240 mg/kg/day or streptozotocin-diabetic mice whenadministered at 60 mg/kg/day (Lord, Atkins, and Bailey, Diabetologia25:108-113, 1983). Metformin caused statistically significant weightloss in KKAy mice treated with 1700 mg/kg/day of the compound, but notwhen lower doses were given (Meglasson, Wilson, Yu, Robinson, Wyse, andde Souza, supra). By comparison, when N-(dihydrazinomethylene)-glycinewas administered to KKAy mice at 100 mg/kg/day it was approximately aseffective as 1700 mg/kg/day of metformin in producing weight loss inthis obese mouse strain (Meglasson, Wilson, Yu, Robinson, Wyse, and deSouza, supra). Metformin has been reported to inhibit non-enzymaticglycosylation of erythrocyte plasma membranes at concentrations of 0.5and 5 micromoles per liter based on its ability to prevent the decreasein the electron paramagnetic resonance spectroscopy order parameters ofplasma membranes incubated with glucose in vitro (Freisleben, Ruckert,Wiernsperger, and Zimmer, Biochemical Pharmacology, 43:1185-1194, 1992).At higher concentrations, 50 and 100 micromoles per liter, metformin hadthe reverse effect and caused a very low order parameter. Hence, whethermetformin could be expected to lessen or aggravate non-enzymaticglycosylation of proteins in diabetic patients would depend on theconcentration of metformin in serum of treated patients. In diabetichumans administered 1 gram of metformin orally, the average Cmax plasmaconcentration is 3.24 micrograms per milliliter (or 25 micromoles perliter) (Tucker, Casey, Phillips, Connor, et al., Br. J. Clin.Pharmacol., 2:235-246, 1981) and, therefore, lies midway between thehighest concentration shown to reduce non-enzymatic glycosylation oferythrocytes and the lowest concentration shown to stimulate theprocess. Based on the published metformin plasma levels in diabeticpatients no conclusion can be drawn as to whether metformin wouldinhibit the non-enzymatic glycosylation of proteins or aggravate theprocess in some manner when administered as a therapy to patients.

General methods for the preparation of the compounds of this inventionare outlined in Schemes 1-4. Specific examples for a number of thesetechniques can be found in the experimental procedures presented in theDescription of the Preferred Embodiment. By using other startingmaterials and reactants the various compounds of the invention may beprepared. The following references discuss procedures relating to thegeneral syntheses of the compounds of this invention.

Scheme 1

Gante, J. Chem. Ber. 1968, 101, 1195. Armarego, W. L. F.; Kobayashi, T.J. Chem. Soc. (C) 1971, 238. Evans, D. A.; Britton, T. C.; Dorow, R. L.;Dellaria, J. F. J. Am. Chem. Soc. 1986, 108, 6395. Evans, D. A.;Britton, T. C.; Dorow, R. L.; Dellaria, J. F. Tetrahedron 1988, 44,5525.

Scheme 3

Gut, J.; Hesoun, D.; Novacek, A. Coll. Czech. Chem. Comm. 1966, 31,2014. Miura, K.; Ikeda, M.; Kondo, T.; Setogawa, K. Chem. Abstr. 1962,56:4767b. Pankaskie, M.; Abdel-Monem, M. M. J. Pharm. Sci. 1980, 69,1000.

Scheme 4

Lee, K.; Kim, S.; Um, H.; Park, H. Synthesis 1989, 638. Reddy, T. I.;Bhawal, B. M.; Rajappa, S. Tetrahedron 1993, 49, 2101.

In Vivo and In Vitro Screening Protocols

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following experimental procedures are specific examples whichdescribe the preparation of a number of compounds of the invention:

EXAMPLE 1

[2-(aminoiminomethyl)hydrazino]-acetic acid

Ethylhydrazinoacetate hydrochloride (7.73 g, 50 mmol) was saponified byrefluxing in 100 mL of 1N NaOH for 2 h. To the hot solution was thenadded 2-methyl-2-thiopseudourea sulfate (6.95 g, 50 mmol) and thesolution was refluxed for an additional 2 h. The mixture wasconcentrated to ˜1/2 volume at which time a white solid precipitated.The solution was cooled and filtered to yield 3.34 g of a white solid.Recrystallization from water afforded 2.41 g (36%) of[2-(aminoiminomethyl)hydrazino]-acetic acid as a highly crystallinewhite solid.

MP: 247-248° C. (dec)

¹ H NMR: (D₂ O) δ 3.40 (s, 2H).

EXAMPLE 2

[2-(aminoiminomethyl)hydrazino]-, monohydrochloride acetic acid

To a stirring solution of [(aminoiminomethyl)hydrazino]-,monohydrochloride, monohydrate acetic acid (10 g, 60 mmol) in methanol(300 ml) was added 10% Pd--C (0.25 g) and the mixture hydrogenated at 30psi overnight. The mixture was filtered and solvent evaporated todryness. The residue was recrystallized from EtOH to afford 4.2 g (42%)title compound as a white solid

(m.p. 163-165° C.).

¹ H NMR (D₂ O) δ 3.68 (s, 2H).

EXAMPLE 3

[2-(aminoiminomethyl)hydrazino]acetic acid phenylmethyl estermonohydrochloride]

HCl (g) was bubbled through a suspension of[2-(aminoiminomethyl)hydrazino]-acetic acid (2.00 g, 15.2 mmol) inbenzyl alcohol (30 mL). The reaction was stirred for about an hour untileverything was in solution. The crude product was precipitated out byadding Et₂ O. This material was recrystallized from MeOH/EtOAc to yield[2-(aminoiminomethyl)hydrazino]acetic acid phenylmethyl estermonohydrochloride (3.20 g, 82%) as a white crystalline solid.

MP: 162-164° C.

¹ H NMR (CD₃ OD): δ 3.69 (s, 2H), 5.24 (s, 2H), 7.34-7.42 (m, 5H).

EXAMPLE 4

α-hydrazinobenzenepropanoic acid

A solution of LDA (50 mL of a 1.5M solution in THF) in 250 mL of dry THFwas cooled to -78° C. To this was added dropwise a solution ofethylhydrocinnamate (12.0 mL, 68.2 mmol) in 250 mL dry THF. The solutionwas stirred at -78° C. for 30 min. A solution of di-tert-butylazodicarboxylate (18.84 g, 81.8 mmol) in 100 mL dry THF was then addeddropwise. After 10 min, the reaction was quenched by the addition of 14mL HOAc and allowing to warm to room temperature. The mixture waspartitioned between Et₂ O and water. The aqueous layer was extractedwith Et₂ O (3×100 mL). The combined organic layers were washed withsaturated aq NaHCO₃ (2×100 mL) and brine (1×100 mL), dried over sodiumsulfate and condensed. The crude product was chromatographed on silica(90/10 hexane/EtOAc) to afford 15.33 g (55%) of the bis-BOC protectedhydrazino ester. The ester was taken up in 200 mL CH₂ Cl₂. To this wasadded 120 mL of trifluoroacetic acid. The mixture was stirred 2 h atroom temperature. After removal of the solvent, the crude product wastaken up in 75 mL of 1N NaOH and refluxed for 2 h. The solution wascooled, extracted with Et₂ O, neutralized, condensed to half volume,cooled and filtered. The resulting brownish solid was stirred in boilingi-PrOH for 5 min to remove colored impurities. Filtration and dryingyielded 3.35 g (27%) of α-hydrazinobenzenepropanoic acid as a whitesolid.

MP: 198-201° C. (dec).

¹ H NMR: (D₂ O) δ 7.41-7.29 (m, 5H), 3.89 (dd, J=7, 6 Hz, 1H), 3.23-3.08(m, 2H).

EXAMPLE 5

α-[2-(aminoiminomethyl)hydrazino]benzenepropanoic acid monohydrate]

A solution of α-hydrazinobenzenepropanoic acid (3.00 g, 16.7 mmol) and2-methyl-2-thiopsuedourea sulfate (2.55 g, 18.3 mmol) in 17 mL 1N NaOHwas heated to reflux for 2 h. The mixture was neutralized with 3N HCland concentrated until precipitation began (ca. 1/2 volume). The crudeproduct was filtered and recrystallized from water to yield 1.81 g (49%)of α[2-(aminoiminomethyl)hydrazino]benzenepropanoic acid monohydrate asa monohydrate.

MP: 127-130° C. (dec).

¹ H NMR: (D₂ O) δ 7.40-7.27 (m, 5H), 3.60 (dd, J=8, 6 Hz, 1H), 3.04 (dd,J=14, 6 Hz, 1H), 2.86 (dd, J=14, 8 Hz, 1H).

2-[2-(Aminoiminomethyl)hydrazino]propanoic acid.

A mixture of 10.0 g (55.4 mmol) 2-[(aminoiminomethyl)hydrazino]propanoicacid hydrochloride (J. Pharmaceut. Sci. 1980, 69, 1000-1004), 1.5 g of10% palladium on carbon, and 300 mL of distilled water was shaken under50 psi hydrogen pressure for 16 h at 25° C. The mixture was filtered. Tothe filtrate was added 75 g of Dowex IR118H hydrogen form stronglyacidic cation exchange resin. The mixture was stirred 1 hour and thenthe mixture was filtered. The resin was washed with three 150 mLportions of distilled water. The combined filtrate and washes werediscarded and the resin was washed with five 200 mL portions of 20%(vol./vol.) pyridine in distilled water. These washes were combined andthe solvent was evaporated at reduced pressure (25° C., 1 torr). Theresulting white powder was dissolved in 30 mL of refluxing distilledwater and the resulting solution was diluted with 90 mL of hot absoluteethanol. The mixture was allowed to cool to 25° C., and after 24 h theprecipitate which formed was collected by filtration. The solid wasdried (20 torr/50° C./24 hours) to give 3.8 g of the title compound as awhite solid,

mp 239-241° C.

EXAMPLE 6

[1-(aminoiminomethyl)hydrazino]acetic acid monohydrobromide

To a stirring suspension of aminoguanidine bicarbonate (100 g, 734 mmolwater (200 ml) was added bromoacetic acid (100 g, 720 mmol). Afterinitial effervescence the homogeneous solution was refluxed overnight,cooled to ambient temperature, and solvent evaporated to dryness. Theresidue was suspended in EtOH (200 ml) and sonicated. The solid wasfiltered to afford 13.6 g (9%) of title compound as a white solid.

(m.p. 163-165° C.).

¹ H NMR (D₂ O) δ 4.25 (s, 2H).

EXAMPLE 7

3-[[imino[(1-methylethylidene)hydrazino]methyl]amino]propanoic acid

β-alanine (6.00 g, 67.5 mmol) was dissolved in 67.5 mL of 1N NaOH. Tothis was added N-amino-S-methylisothiourea hydroiodide (15.69 g, 67.5mmol). The mixture was heated to reflux for 1.5 h. The solvent wasremoved. The crude product was taken up in ca. 50 mL water and 50 mL ofacetone was added. Removal of the solvent afforded an orange solid whichwas chromatographed on silica (80/20 CHCl₃ /MeOH then 60/40 CHCl₃ /MeOH)to yield 5.88 g (47%) of3-[[imino[(1-methylethylidene)hydrazino]methyl]amino]propanoic acid as apale orange solid.

MP: ˜125° C. (dec).

¹ H NMR: (D₂ O) δ 3.36 (t, J=6 Hz, 2H), 2.35 (t, J=6 Hz, 2H), 1.87 (s,3H), 1.80 (s, 3H).

EXAMPLE 8

N-(hydrazinoiminomethyl)-β-alanine

3-[[imino[(1-methylethylidene)hydrazino]methyl]amino]propanoic acid(5.88 g, 31.61 mmol) was dissolved in 125 mL water and heated to 60° C.for 72 h. The solvent was evaporated and the product was stirred in a4:1 mixture of EtOH and MeOH. The resulting pale orange precipitate wasfiltered, washed with ethanol and dried to yield 3.16 g (68%) ofN-(hydrazinoiminomethyl)-β-alanine as a pale orange solid.

MP: 177-179° C.

¹ H NMR: (D₂ O) δ 3.39 (t, J=6 Hz, 2H), 2.42 (t, J=6 Hz, 2H).

EXAMPLE 9

N-(dihydrazinomethylene)-1-alanine

To a suspension of L-alanine (10.0 g, 0.11 mol) and triethylamine (33.5mL, 0.24 mol) in EtOH (90 ml) and H₂ O (6 mL) was added carbon disulfide(7.2 mL, 0.12 mol). After stirring overnight, methyl iodide (7.5 mL,0.12 mol) was added to the yellow solution. The mixture was stirred for1 h and concentrated to a slurry. The residue was dissolved in H₂ O (25mL), and conc. HCl was added until acidic. The mixture was extractedwith Et₂ O (3×100 mL), and the organic phase was dried (MgSO₄) andconcentrated to provide 18.4 g (93%) of the correspondingdithiocarbamate as a pale yellow solid of good purity. A analyticallypure sample was obtained by recrystallization from Et₂ O/hexane.

m.p. 90-92;

¹ H NMR (D₂ O) δ 4.89 (q, J=7 Hz, 1 H), 2.59 (s, 3 H), 1.52 (d, J=7 Hz,3 H).

To a solution of the dithiocarbamate (5.0 g, 28 mmol) in methylenechloride (50 mL) at 0° C. was added methyl trifluoromethanesulfonate(3.5 mL, 31 mmol). The mixture was warmed to room temperature andstirred for 20 h. The mixture was concentrated under reduced pressure toa colorless oil. The resulting oil was dissolved in H₂ O (5 mL), and 1.0M NaOH (28 mmol) was added. The mixture was extracted with EtOAc (3×100mL), and the organic phase was dried (MgSO₄). After filtration, thesolvent was removed in vacuo to provide a thick viscous oil. The oil wasdissolved in absolute EtOH (25 mL), and anhydrous hydrazine (4.4 mL,0.14 mol) was added. The mixture was stirred for 1.5 h, and the solid(2.5 g) which formed was collected by filtration. The white powder wasfurther purified by crystallization from H₂ O/IPA to give 2.2 g (49%) ofthe diaminoguanidine as a white powder.

m.p. 174-176 (dec.)

¹ H NMR (D₂ O) δ 3.69 (q, J=7 Hz, 1 H), 1.20 (d, J=7 Hz, 3 H).

EXAMPLE 10

N-(dihydrazinomethylene)-β-alanine

By a procedure analogous to that employed forN-(dihydrazinomethylene)-1-alanine, β-alanine was converted toN-(dihydrazinomethylene)-β-alanine.

(m.p. 192° C., dec.).

¹ H NMR (D₂ O) 3.40 (t, 2H, J=7 Hz), 2.48 (t, 2H, J=7 Hz).

EXAMPLE 11

N-(dihydrazinomethylene)-glycine

A solution of methylated thiocarbohydrazide (25.0 g, 101 mmol) andglycine (6.314 g, 83.98 mmol) in water (50 mL) and 12.5 N NaOH (8.89 mL,111 mmol) was stirred under nitrogen at 75-80° C. for 3 hrs. Thesolution was chilled in ice while still under nitrogen before theportionwise addition of abs. ethanol (550 mL in 50 mL portions),stirring between each addn until pptn was complete. The mixture was thenstirred for 15 min. at 0° C. before filtering. The collected solid waswashed thoroughly with abs. ethanol. Drying gave a lt. pink powder (8.04g). The crude solid was dissolved in water (30 mL), filtered to removesome fine insoluble material, and then diluted to a volume of 250 mLwith abs. ethanol. Precipitation began almost immediately and wasaccelerated by sonication for a few seconds. After standing at room tempfor 10 min, the mixture was filtered, giving a pale rose powder (5.25 g,42%.

m.p. 200° C., dec.).

¹ H NMR (D² O) 3.78 (s).

EXAMPLE 12

[2-(hydrazinoiminomethyl)hydrazino]acetic acid

Ethylhydrazinoacetate hydrochloride (9.28 g, 60 mmol) was saponified byrefluxing in 120 mL of 1N NaOH for 2 h. To the hot solution was thenadded N-amino-S-methylisothiourea hydroiodide (13.98 g, 60 mmol) and thesolution was refluxed for an additional 2 h. The solvent was removed.The crude product was dissolved in methanol and filtered to remove theNaCl. The filtrate was condensed and dried by high vac. The residue wasthen stirred with 150 mL MeOH overnight. The resulting white solid wasfiltered. This solid was then refluxed in 100 mL MeOH for 2 h to removeany impurities. The mixture was then cooled and filtered. The resultingsolid was dried in vacuo to yield 2.14 g (24%) of[2-(hydrazinoiminomethyl)hydrazino]acetic acid as an off-white solid.

MP: 201-203° C.(dec).

¹ H NMR: (D₂ O) δ 3.39 (s, 2H).

EXAMPLE 13

N-(dihydrazinomethylene)-d-alanine

To a suspension of D-alanine (1.8 g, 20 mmol) and triethylamine (6.1 mL,44 mmol) in EtOH (15 ml) and H₂ O (1 mL) was added carbon disulfide (1.3mL, 22 mmol). After stirring overnight, methyl iodide (1.4 mL, 22 mmol)was added to the yellow solution. The mixture was stirred for 1 h andconcentrated to a slurry. The residue was dissolved in H₂ O, and conc.HCl was added until acidic. The mixture was extracted with methylt-butyl ether (3×50 mL), and the organic phase was dried (MgSO₄) andconcentrated to provide a yellow oil, which with sonication and theaddition of a small amount of hexane solidified. Upon further drying,2.9 g of a yellow solid was obtained. The product was further purifiedby recrystallization (Et₂ O/hexane) to give 1.67 g (47%) of the compoundidentified as compound A of Table 9 as a cream solid.

m.p. 89-91° C.;

¹ H NMR (D₂ O) δ 4.67 (m, 1 H), 2.39 (s, 3 H), 1.32 (d, J=7.0 Hz, 3 H).

To a solution of the dithiocarbamate of Compound A of Table 9 (15.1 g,84.3 mmol) in methylene chloride (170 mL) at 0° C. was added methyltrifluoromethanesulfonate (10.5 mL, 92.7 mmol). The mixture was warmedto room temperature and stirred for 20 h. The mixture was concentratedunder reduced pressure to a colorless oil. The resulting oil wasdissolved in H₂ O (40 mL), and 1.0 M NaOH (84.3 mmol) was added. Themixture was extracted with EtOAc (3×200 mL), and the organic phase wasdried (MgSO₄). After filtration, the solvent was removed in vacuo toprovide a thick viscous oil. The oil was dissolved in absolute EtOH (85mL), and anhydrous hydrazine (13.2 mL, 0.42 mol) was added. The mixturewas stirred for 1.5 h, and the solid (7.5 g) which formed was collectedby filtration. The white powder was further purified by crystallizationfrom H₂ O/IPA to give 6.48 g (48%) of the title compound as a whitepowder.

m.p. 175-177° C.;

H NMR (D₂ O) δ 3.69 (q, J=7 Hz, 1 H), 1.20 (d, J=7 Hz, 3 H).

EXAMPLE 14

N-(dihydrazinomethylene)-valine

To a suspension of L-valine (5.0 g, 42.7 mmol) and triethylamine (13.1mL, 93.9 mmol) in EtOH (30 ml) and H₂ O (2 mL) was added carbondisulfide (2.8 mL, 47.0 mmol). After stirring overnight, methyl iodide(2.9 mL, 47.0 mmol) was added to the yellow solution. The mixture wasstirred for 2 h and concentrated to a slurry. The residue was dissolvedin H₂ O (10 mL), and conc. HCl was added until acidic. The mixture wasextracted with Et₂ O (3×100 mL), and the organic phase was dried (MgSO₄)and concentrated to provide a yellow oil which after seeding gave ayellow solid. The solid was suspended in hexane and filtered to yield7.7 g of Compound B of Table 9 as an off-white solid. The filtrate wascooled to 0° C. to yield a second crop of 0.27 g of Compound B of Table9 (7.97 g total, 90%) as a white solid.

m.p. 76-78° C.;

¹ H NMR (CDCl₃) δ 5.30 (m, 1 H), 2.40 (m, 1 H), 1.08 (d, J=7.0 Hz, 3 H),1.04 (d, J=7.0 Hz, 3 H).

To a solution of Compound B of Table 9 (8.0 g, 38.6 mmol) in methylenechloride (60 mL) at 0° C. was added methyl trifluoromethanesulfonate(4.8 mL, 42.5 mmol). The mixture was warmed to room temperature andstirred for 20 h. The mixture was concentrated under reduced pressure toa colorless oil. The resulting oil was dissolved in H₂ O (10 mL), and1.0 M NaOH (38.6 mL) was added. The mixture was extracted with EtOAc(3×100 mL), and the organic phase was dried (MgSO₄). After filtration,the solvent was removed in vacuo to provide a thick viscous oil. The oilwas dissolved in isopropyl alcohol (150 mL), and hydrazine monohyrate(9.4 mL, 0.19 mol) was added. The mixture was stirred for 2 h, and THFwas added which resulted in a more filterable solid. Filtration provided2.4 g (33%) of the title compound as a slightly hygroscopic white solid.

m.p. 112-116° C.;

¹ H NMR (D₂ O) δ 3.70 (d, J=5.0 Hz, 1 H), 2.20 (m, 1 H), 0.97 (d, J=7.0Hz, 3 H), 0.94 (d, J=7.0 Hz, 3 H).

EXAMPLE 15

[1-(aminohydrazonomethyl)hydrazino]acetic acid (Please Refer to Scheme5)

Preparation of 9

To a stirring suspension of ethyl hydrazinoacetate hydrochloride (5.0 g,32.34 mmol) and N-methyl morpholine (3.26 g, 32.34 mmol) at 0° C. wasadded solid N-(Benzyloxycarbonyloxy)succinimide (8.06 g, 32.34 mmol).The mixture was allowed to warm to ambient temperature overnight and thesolvent removed in vacuo. The residue was suspended between EtOAc/H₂ O,the layers shaken, the organics separated and dried over Na₂ SO₄. Thesolvent was removed and the residue chomatographed via SiO₂ flashchromatography (eluant 4:1 hexane/EtOAc) to afford 5.7 g (70%) titlecompound as a white solid. m.p. 95-97° C. The residue in subsequentreactions was purified by recrystallization from EtOAc/hexane to affordtitle compound in slightly lower yield.

¹ H NMR (CDCl₃) δ 1.27 (t, J=7 Hz, 3 H), 3.66 (s, 2 H), 4.19 (q, J=7 Hz,2 H), 5.13 (s, 2 H), 6.77 (brs, 1 H), 7.33 (m, 5 H).

Preparation of 10

To a stirring suspension of Preparation 9 (3.0 g, 11.89 mmol) in EtOH(30 ml) at ambient temperature, was added aqueous NaOH (1N, 11.89 ml).To the mixture was added additional H₂ O (10 ml) and stirred for 1 hr(The mixture became a homogeneous solution and then a solidprecipitated). Aqueous HCl (1 N, 11.89 ml) was then added, the ethanolremoved in vacuo and the aqueous extracted with EtOAc (2×100 ml). Theorganic layers were combined, dried over Na₂ SO₄, and the solventremoved to afford 2.31 g (87%) title compound as white solid.

m.p. 131-133° C.

¹ H NMR (CD₃ OD) δ 3.59 (s, 2 H), 5.15 (s, 2 H), 7.37 (m, 5 H).

Preparation of 11

To a stirring suspension of Preparation 10 (25.44 g, 112.7 mmol) inEtOAc (500 ml) was added trimethylsilyl isothiocyanate (14.79 g, 112.7mmol) and the mixture was heated at gentle reflux (80° C.) overnight.The resulting solution was cooled to ambient temperature and washed withH₂ O (2×100 ml). The organic layer was separated, dried over Na₂ SO₄,and the solvent evaporated to dryness. The oily residue was dissolved inCH₂ Cl₂ and allowed to stand at ambient temperature for 3 min in whichtime a solid forms. The solid was filtered, washed with CH₂ Cl₂ (100 ml)and dried in vacuo. The solid was slurried in hot EtOAc (300 ml) todissolve any sulphur related by-products and triturated with hexane (200ml) to afford 17.1 g title compound (53%) as a white solid.

m.p. 148-149° C.

¹ H NMR (CD₃ OD) δ 5.20 (s, 2 H), 7.30 (m, 5 H) remaining CH2 notobservable.

Preparation of 12

To a stirring solution of Preparation 11 (5.0 g, 17.64 mmol) in EtOH(150 ml) at ambient temperature was added methyl iodide (2.73 g, 19.41mmol) and the resulting solution stirred overnight. The solvent wasremoved in vacuo to afford 7.50 g (quant) title compound as a yellowfoam.

¹ H NMR (CD₃ OD) δ 2.69 (brs, 0.6 H), 2.84 (brs, 0.4H), 4.40-4.70 (m,2H), 5.31 (brs, 2H), 7.46 (m, 5H).

Preparation of 13

To a vigorously stirring solution of Preparation 12 (25.5 g, 60 mmol) inH₂ O (100 ml) at ambient temperature was added hydrazine hydrate (6.06g, 120 mmol) slowly until 1/2 had been added. H₂ O (10 ml) was added tothe solid mass which had formed and the solids broken up mechanicallywith a spatula. The remaining hydrazine was then added and the solutionvigorously stirred for 1 hour. The heterogeneous mixture was sonicatedand stirring continued until a thick mass had formed. EtOH (50 ml) wasadded, the solid filtered, washed with EtOH and dried in vacuo to afford9.24 g (55%) title compound as a white solid.

m.p. 168-170° C.

¹ H NMR (D₂ O) δ 3.86 (brs, 1 H), 4.21 (brs, 1 H), 5.17 (s, 2 H), 7.39(s, 5 H).

[1-(hydrazinoiminomethyl)hydrazino]acetic acid

To a solution of Preparation 13 (9.20 g, 32.71 mmol) in MeOH/H₂ O (400ml, ˜2:1 v/v) was added 10% Pd--C (1.0 g) and the mixture hydrogenatedat 30 psi for 4 hours. The catalyst was filtered through diatomaceousearth and 10% Pd--C (1.0 g) was again added. The mixture washydrogenated at 30 psi for 2.5 hours and determined to be complete byTLC (eluant 85:14:1 CH₂ Cl₂ /MeOH/HCO₂ H). The mixture was filteredthrough diatomaceous earth and solvent removed to ˜50 ml at which time asolid precipitated. The solid was filtered, washed with a minimum amountof H₂ O and dried in vacuo to afford 3.60 g (75%) title compound as anoff white solid. m.p. 196-198° C. A second crop was obtained byconcentrating the filtrate until a solid formed. Filtration afforded0.90 g (19%, total yield: 94%) additional material having identicalmelting point.

¹ H NMR (D₂ O) δ 4.06 (s, 2 H).

BIOLOGICAL TESTING

Compounds of the present invention were tested for their ability toreduce blood glucose and body weight as follows:

KKAy mice are rodent models of NIDDM and obesity (Chang, Wyse, Copeland,Peterson, and Ledbetter, 1986). A pre-treatment blood sample wasobtained from the retro-orbital sinus and the mice arranged in groups of6 so that the mean pre-treatment blood glucose level was the same onaverage in all groups. Test compounds were admixed in the chow at aconcentration of 0.5% and the mice were allowed to consume the diet adlibitum. Control mice received unsupplemented chow. On Day 0, the micewere weighed and provided control chow or chow supplemented with testcompounds. After 3 days of consuming control chow or chow supplementedwith test compounds, a blood sample was obtained for determination ofthe glucose concentration and the animals were weighed for determinationof weight loss. Food consumption was measured by weighing the foodprovided at the beginning of the study and the food residue at the endof the study. Food consumption was calculated by subtracting the weightof the residue from the weight of the food provided. Drug intake wascalculated by multiplying food consumption by 0.5%. Using this methoddrug intake was determined to be approximately 500 mg/kg/day. Bloodglucose data are expressed as the average blood glucose concentration inthe test group divided by the average blood glucose level in the controlgroup (treatment/control or T/C). Compounds resulting in T/C valuesequal to or less than 0.90 are considered to be activeanti-hyperglycemic agents. Weigh loss data are expressed as percentchange in body weight. Compounds resulting in a decrease of 1% or moreless than control in body weight over three days are considered to beactive anti-obesity agents.

                                      TABLE 1                                     __________________________________________________________________________    Preferred Compounds of the Invention                                          __________________________________________________________________________                          Acetic acid, [2-(aminoiminomethyl)-  hydrazino]-                              -                                                                             Acetic acid, [2-(aminoiminomethyl)-  hydrazino]-mono                        hydrochloride                                                -                                                                                                Glycine, N-(hydrazinoiminomethyl)-                         -                                                                                                Glycine, N-(hydrazinoiminomethyl)-,  hydrochloride                          (2:1)                                                        -                                                                                                N-(Dihydrazinomethylene)-glycine                           -                                                                                                Acetic acid, [2-(hydrazinoiminomethyl-  hydrazino]-        -                                                                                                [1-(hydrazinoiminomethyl)hydrazino]acetic  acid         __________________________________________________________________________

                                      TABLE 2                                     __________________________________________________________________________    Related Inactive Compounds Which are not Claimed                              __________________________________________________________________________                          Acetic acid, [(aminoiminomethyl)-  methylhydrazono],                         sulfate (2:1)                                               -                                                                                                Acetic acid, [[imino(nitroamino)-  methyl]hydrazono]                        -                                                            -                                                                                                Acetic acid, [[imino(methylamino)-  methyl]hydrazono                        ]-                                                           -                                                                                                #STR14##                                                   -                                                                                               ##STR15##                                                __________________________________________________________________________

                                      TABLE 3                                     __________________________________________________________________________    Inactive Exceptions to the Generic Scope                                      __________________________________________________________________________                            Propanoic acid, 3-                                                          [1-(amino-  iminomethyl)hydrazino]-                        -                                                                                                  #STR17##                                                 -                                                                                                  Butanoic acid, 2-[2-(amino-  inimomethyl)hydrazino                          ]-                                                         -                                                                                                  #STR19##                                                 -                                                                                                  Butanoic acid, 4-[(hydrazino-  iminomethyl)amino]-    __________________________________________________________________________

                                      TABLE 4                                     __________________________________________________________________________    Specifically Claimed Compounds of the Invention                               __________________________________________________________________________                                  Acetic acid, [2-(aminoiminomethyl)-                                         hydrazino]-                                          -                                                                                                        Acetic acid, [2-(aminoiminomethyl)-                                         hydrazino]-, monohydrochloride                       -                                                                                                        Acetic acid, [2-(aminoiminomethyl)-                                         hydrazino]-, phenylmethyl ester,  monohydrochl                                oride                                                -                                                                                                        Benzenepropanoic acid, alpha[2-  (aminoimino                                methyl)hydrazino],   monohydrate                     -                                                                                                        #STR25##                                           -                                                                                                        Acetic acid, [1-(aminoiminomethyl)-                                         hydrazino]-, monohydrobromide                        -                                                                                                        β-Alanine, N-(hydrazinoiminomethyl)-                                     -                                                                             N-(Dihydrazinomethylene)-glycine                   -                                                                                                        Acetic acid, [2-(hydrazinoiminomethyl)-                                     hydrazino]-                                          -                                                                                                        β-Alanine, N-(dihydrazinomethylene)-                                     -                                                                             L-Alanine, N-(dihydrazinomethylene)-                                          -                                                                             N-(dihydrazinomethylene)-d-alanine                 -                                                                                                        N-(dihydrazinomethylene)-valine                    -                                                                                                        [1-(aminohydrazonomethyl)hydrazino]acetic                                   acid                                              __________________________________________________________________________

                                      TABLE 5                                     __________________________________________________________________________    Known Compounds Claimed for Treatment of NIDDM                                __________________________________________________________________________                             Acetic acid,  [(aminoiminomethyl)hydrazono]-,                               monohydrochloride, monohydrate                            -                                                                                                   [Propanoic acid, 2-[(amino-  iminomethyl)hydrazon                           o]-,  monohydrochloride                                   -                                                                                                   Butanoic acid, 2-[(amino-  iminomethyl)hydrazono]                           -,  monohydrochloride                                     -                                                                                                   Glycine, N-(hydrazinoiminomethyl)-                      -                                                                                                   Glycine, N-(hydrazinoiminomethyl)-,  hydrochlorid                           e (2:1)                                                   -                                                                                                   Benzenepropanoic acid, α-[(amino-                                     iminomethyl)hydrazono]-                                   -                                                                                                   Octanoic acid, 2-[(aminoiminomethyl)-  hydrazono]                           -, monohydrochloride                                   __________________________________________________________________________

Table 6

Dose-Response for Reduction in Hyperglycemia and Obesity in KKAy mice byOral Administration of N-(dihydrazinomethylene)-glycine

KKAy mice were treated with N-(dihydrazinomethylene)-glycine asdescribed above except that the compound was admixed in the chow at0.03, 0.06, 0.10, 0.20, 0.30, and 0.40% so as to deliver daily doses ofapproximately 30, 60, 100, 200, 300, and 400 mg/kg. Control micereceived unsupplemented chow. For comparison toN-(dihydrazinomethylene)-glycine, 3-guanidinopropionic acid (3-GPA) wasadministered as a 0.50% admixture in the chow (approximate dose, 500mg/kg/day). Data are shown as the percent change in blood glucoseconcentration and body weight on Day 3 compared to Day 1 of the study.Means±S.E.M. for n=6 mice/group. Statistical significance was determinedby analysis of variance using JMP 3.0.2 software (SAS Institute). *,P<0.05 vs. Nil; , significantly less than 3-GPA (P<0.05).

    ______________________________________                                                         % Change    % Change                                           Addition Blood Glucose Body Weight                                          ______________________________________                                        Nil               -5.8 ± 7.1                                                                            -0.71 ± 0.65                                    N-(dihydrazinomethylene)-glycine                                              0.03% -13.5 ± 10.5 -0.92 ± 0.35                                         N-(dihydrazinomethylene)-glycine                                              0.06% -34.9 ± 17.1* -1.51 ± 2.11                                        N-(dihydrazinomethylene)-glycine                                              0.10% -45.2 ± 6.4* -4.04 ± 0.76*                                        N-(dihydrazinomethylene)-glycine                                              0.2% -69.9 ± 3.2*,  -8.22 ± 1.05*                                       N-(dihydrazinomethylene)-glycine                                              0.3% -70.4 ± 1.5*,  -9.94 ± 1.62*,                                      N-(dihydrazinomethylene)-glycine                                              0.4% -70.3 ± 3.9*,  -10.3 ± 0.97*,                                      0.5% 3-GPA -38.4 ± 4.4*  -5.4 ± 0.81*                                 ______________________________________                                         *, P < 0.05 vs. Nil                                                            , significantly less than 3GPA (P < 0.05)                               

Table 7

Improvement of Intraperitoneal Glucose Tolerance

Glucose tolerance was measured in non-diabetic C57BL mice and diabeticKKAy mice. The animals were dosed by oral gavage with distilled water(Control) or 100 mg/kg of N-(dihydrazinomethylene)-glycine then fastedfor 16-17 hours. Blood samples for glucose determination were obtainedfrom the retro-orbital sinus. Samples were obtained immediately prior toadministration of 2 g/kg glucose I.P. (Time=0) and 30, 60, and 90minutes after the injection. Blood glucose was determined using aglucose autoanalyzer. The data are expressed as means±S.E.M. for 5-6mice per group. Statistical significance was determined by analysis ofvariance using JMP 3.0.2 software (SAS Institute). *, P<0.05 vs.Control.

    ______________________________________                                        Mouse Strain                                                                           Group       Time (min.)                                                                             Blood Glucose (mg/dl)                          ______________________________________                                        C57BL    Control      0        143 ± 8                                         30 233 ± 14                                                                60 240 ± 8                                                                 90 226 ± 9                                                                N-(dihydrazino-  0 114 ± 9*                                                methylene)- 30 174 ± 17*                                                   glycine 60 153 ± 7*                                                         90 161 ± 19*                                                             KKAy Control  0 188 ± 43                                                     30 487 ± 10                                                                60 469 ± 20                                                                90 486 ± 26                                                               N-(dihydrazino-  0 115 ± 16                                                methylene)-  (P = 0.12                                                        glycine  vs. Control)                                                          30 383 ± 38*                                                               60 396 ± 63                                                                90 392 ± 67                                                            ______________________________________                                    

Table 8

Inhibition of Non-Enzymatic Glycosylation of Protein

Non-enzymatic glycosylation of protein was measured using establishedmethods (Dolhofer and Wieland, 1979; Khatami, Suldan, David, Li, andRockey, 1988). The incorporation of 100 mM [14C]-D-glucose into humanserum albumin was determined by dissolving human serum albumin (SigmaChemical Co.), [14C]-glucose, and glucose in a physiological salinesolution and incubating at 37° C. for 8 days. Test compounds were addedto the solution at 19.1 mM. Glycosylation of albumin was determined byprecipitating the protein with 1 volume 12% trichloroacetic acid,centrifuging, and washing the pellet twice with 6% trichloroacetic acidwith centrifugation following each wash. The washed pellet wassolubilized, scintillant added and the incorporation of radiolabelledglucose determined by liquid scintillation counting. The data areexpressed as the percent of [14C]-glucose incorporated into albumin(mean of 2 measurements). Statistical significance was determined byanalysis of variance using JMP 3.0.2 software (SAS Institute).

    ______________________________________                                        Substance Added % [14C-glucose] Incorporated                                  ______________________________________                                        Control (Nil)   1.50                                                            Aminoguanidine 0.96                                                            (P < 0.05                                                                     vs. Control)                                                                 3-Guanidinopropionic Acid 1.52                                                N-(dihydrazinomethylene)- 0.81                                                glycine (P < 0.05                                                              vs. Control)                                                                 N-(hydrazinoiminomethyl)- 1.21                                                glycine (P < 0.05                                                              vs. Control)                                                                 monohydrochloride 1.29                                                        acetic acid (P < 0.10                                                          vs. Control)                                                               ______________________________________                                    

                  TABLE 9                                                         ______________________________________                                        Intermediate Compounds                                                        ______________________________________                                                              Compound A                                                 -                                                                                                Compound B                                              ______________________________________                                         ##STR44##

What is claimed is:
 1. A method of treating obesity comprisingadministering to a patient in need thereof an effective amount of acompound of the formula III

    HN═C(NH.sub.2)--NH--N═CR.sup.3 --COOH

wherein R³ is H, methyl, ethyl, benzyl or n-hexyl.
 2. A method oftreating obesity comprising administering to a patient in need thereofan effective amount of a compound that is selected from the groupconsisting of:[(aminoiminomethyl)hydrazino]acetic acid,monohydrochloride; 2-[(aminoiminomethyl)hydrazino]propanoic acid,monohydrochloride; 2-[(aminoiminomethyl)hydrazino]butanoic acid,monohydrochloride; α-[(aminoiminomethyl)hydrazino]benezenepropanoicacid; and 2-[(aminoiminomethyl)hydrazino]octanoic acid.
 3. A method oftreating obesity comprising administering to a patient in need thereofan effective amount of a compound that is N-(hydrazinoimomethyl)glycineor its hydrochloride (2:1).
 4. The method of claims 1, 2 or 3, whereinsaid compound is administered to the patient by an oral, parenteral,intranasal, buccal, sublingual, intrarectal, or transdermal route ofadministration.
 5. The method of claims 1, 2 or 3, wherein said compoundis administered to the patient by an oral route.
 6. The method of claims1, 2 or 3, wherein said compound is administered to the patient in anamount of between 0.1 and 100 mg/kg body weight daily.
 7. The method ofclaim 1, 2 or 3, wherein said compound is administered to the patient inan amount of 1-50 mg/kg/day.
 8. The method of claim 4, wherein saidcompound is administered to the patient in an amount of between 0.1 and100 mg/kg body weight daily.
 9. The method of claim 5, wherein saidcompound is administered to the patient in an amount of between 0.1 and100 mg/kg body weight daily.
 10. The method of claim 4, wherein saidcompound is administered to the patient in an amount of 1-50 mg/kg/day.11. The method of claim 5, wherein said compound is administered to thepatient in an amount of 1-50 mg/kg/day.